Clonagem e caracteriza????o da express??o transiente do gene codificador do pept??deo antimicrobiano Cp-tionina II em plantas de Nicotiana benthamiana

• Autor(a) principal: Leite, Michel Lopes
• Data de Publicação: 2019
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações da UCB
• Texto Completo: https://bdtd.ucb.br:8443/jspui/handle/tede/2561
• topic: Magnifection; Transient recombinant biosynthesis; Tobacco Mosaic Virus; Plant defensin.; Defensina vegetal; V??rus do Mosaico do Tabaco; Bioss??ntese recombinante transiente; CNPQ::CIENCIAS BIOLOGICAS

The number of infections caused by microorganism???s resistant to conventionally used antibiotics has increased dramatically in the last decades and it is estimated that by 2050 this number will be of 10 million cases per year worldwide. Therefore, the bioprospection of new molecules is the epicenter of researches in regard to the treatment of pathogens associated to them. Antimicrobial peptides have been considered as promising molecules to be used, alone or combined to antibiotics, to treat super-resistant microorganisms. However, these molecules are produced at low concentration in their source organisms, necessitating strategies to potentiate their production. The heterologous expression of genes in plants by the MagnifectionTM method is shown to be a viable alternative due to the production of the recombinant peptide occurs with a short period of time. In addition, plants are advantageous because of the absence of pathogens common to humans, as well as being able to make necessary post-translational modifications to some peptides, as in the case of defensin Cp-thionin II, which has four disulfide bonds. This defensin was initially isolated from cowpea bean and showed activity against bacteria and fungi, making it a potential molecule for the treatment of resistant microorganisms. Thus, the aim of this work was to produce heterologous recombinant defensin Cp-thionin II in a plant system using the MagnifectionTM method. Data obtained suggest that the molecular cloning of the insert into the expression vector was made correctly, as well as that the heterologous expression of the gene encoding Cp-thionin II was successful. After extraction, the recombinant defensin represented 31,8% of the total soluble proteins of N. benthamiana plants.